Whole-cell extracts (in 50 mM Tris [pH 7.4], 150 mM NaCl, 10% glycerol, 0.5% Triton X-100, and protease inhibitor; Roche) were electrophoresed in polyacrylamide gels and transferred onto nitrocellulose membranes. Primary antibodies were incubated overnight, and secondary antibodies were incubated for 1 h at room temperature. Antibody incubation was performed in 3% BSA in TBS containing 0.1% Tween 20. Anti-α LN 13 (2765S) and anti-GAPDH (9,484) antibodies were obtained from Cell Signaling and used at 1:2,000 and 1:1,000 dilutions, respectively. Anti-SIRT6 (2590S) and anti-SIRT6 (62,739) antibodies were obtained from Cell Signaling and Abcam, respectively, and used at 1:250 dilutions. IRDye 680LT-conjugated goat anti-mouse IgG (H + L; 926–68020) and IRDye 800CW-conjugated goat anti-rabbit IgG (H + L; 926–32211) secondary antibodies were obtained from LI-COR Bioscience and used at a dilution of 1:5,000.
Anti sirt6 2590s
Manufactured by Cell Signaling Technology
Anti-SIRT6 (2590S) is a primary antibody product offered by Cell Signaling Technology. It is designed to detect SIRT6 protein expression.
Automatically generated - may contain errors
Lab products found in correlation
Protease inhibitor, by Roche (1 mentions)
Anti flag, by Merck Group (1 mentions)
Ab191606, by Abcam (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Anti glycogen synthase kinase 3 beta gsk3β, by Cell Signaling Technology (1 mentions)
Anti gst sc 138, by Santa Cruz Biotechnology (1 mentions)
2 protocols using anti sirt6 2590s
1
Western Blot Analysis of SIRT6 Protein
2
Investigating Protein Expression in Colon Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Human colon cancer HCT116 and LoVo cells were harvested after treatment, the total or NP40 protein was extracted, and protein expression was detected by Western blotting as previously described with minor modifications [35] (link). Equal amounts of proteins were size fractionated by 9% to 15% sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. anti-SIRT6 (2590S, Cell Signaling, Danvers, MA), anti-PKCζ (sc-17781, Santa Cruz, CA), anti-PI3K (ab191606, abcam, Cambridge, MA), anti–serine/threonine kinase 1 (AKT) (9272, Cell Signaling, Danvers, MA),anti–glycogen synthase kinase-3 beta (GSK3β) (9315, Cell Signaling, Danvers, MA), anti–serine/threonine protein phosphatase 2A (PP2A) (ab3210, abcam, Cambridge, MA), anti-phospho-(Ser/Thr) (9631, Cell Signaling, Danvers, MA), anti-Flag (F1804, Sigma Aldrich), anti-His (PM032, MBL), anti-GST (sc-138, Santa Cruz, CA), anti-MYC (M047-3, MBL, Japan), anti–α-tubulin (BE0031, EASYBIO, Beijing, China), and anti–β-actin (4967, Cell Signaling, Danvers, MA) were used, and the blots were developed using an enhanced chemiluminescence kit (Amersham Corp.).
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